Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli
The GDF-11 protein has a positive effect on animals and humans in inflammatory and infectious processes, regeneration and rejuvenation, its high expression is observed in many organs and tissues. The relevance of the problem of oral use of many proteins and peptides, including the GDF-11 protein, as...
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| Format: | Article |
| Language: | Russian |
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Murmansk State Technical University
2025-06-01
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| Series: | Vestnik MGTU |
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| Online Access: | https://vestnik.mauniver.ru/show-eng.shtml?art=2271 |
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| author | Valieva Sh. S. Tikhonov S. L. Tikhonova N. V. Timofeeva M. S. Shikhalev S. V. |
| author_facet | Valieva Sh. S. Tikhonov S. L. Tikhonova N. V. Timofeeva M. S. Shikhalev S. V. |
| author_sort | Valieva Sh. S. |
| collection | DOAJ |
| description | The GDF-11 protein has a positive effect on animals and humans in inflammatory and infectious processes, regeneration and rejuvenation, its high expression is observed in many organs and tissues. The relevance of the problem of oral use of many proteins and peptides, including the GDF-11 protein, as drugs and in specialized and functional products is noted. The aim of the research is to develop a proteolysis-resistant peptide sequence of the GDF-11 protein with virtual screening of its toxicity, allergenicity, uniqueness with subsequent synthesis of the corresponding plasmid for protein expression in E. coli. The GDF-11 protein, pET-25b(+) plasmids for expression in E. coli with the peptide sequence CTVDСFFECAFGСWDС have been used as objects of study. The GDF-11 protein gene with the sequence CTVDСFFECAFGСWDС has been synthesized from oligonucleotides using the cyclic assembly method. The obtained plasmid sequence encoding the new GDF-11 protein has been treated with restriction endonucleases BamHI and XhoI and cloned into the pET-25b(+) vector for E. coli expression. The clones have been sequenced by the Sanger method. The distribution and identification of primers in the obtained sequence have been studied by the MALDI-TOF MS Ultraflex method. A new unique peptide sequence CTVDСFFECAFGСWDС with cyclization at the C1-C4, C1-C2, C1-C3 bonds of the GDF-11 protein is developed, which allows increasing its resistance to proteolysis and, accordingly, opens up the possibility of oral use in prophylactic food products. Based on virtual screening, it is established that the new peptide chain CTVDСFFECAFGСWDС of the GDF-11 protein is not toxic to the cardiovascular and nervous systems, is not hepatotoxic, does not have cytotoxicity, does not have allergenicity and is characterized by high plasma clearance and an average half-life. |
| format | Article |
| id | doaj-art-893d7f04811f4ec7b77532f235b49dbf |
| institution | Kabale University |
| issn | 1560-9278 1997-4736 |
| language | Russian |
| publishDate | 2025-06-01 |
| publisher | Murmansk State Technical University |
| record_format | Article |
| series | Vestnik MGTU |
| spelling | doaj-art-893d7f04811f4ec7b77532f235b49dbf2025-08-20T03:29:23ZrusMurmansk State Technical UniversityVestnik MGTU1560-92781997-47362025-06-0128214315010.21443/1560-9278-2025-28-2-143-150Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coliValieva Sh. S.0 Tikhonov S. L.1Tikhonova N. V.2Timofeeva M. S.3Shikhalev S. V.4South Ural State Agrarian UniversityUral State Agrarian UniversityUral State Agrarian UniversityUral State Agrarian UniversityUral State Agrarian UniversityThe GDF-11 protein has a positive effect on animals and humans in inflammatory and infectious processes, regeneration and rejuvenation, its high expression is observed in many organs and tissues. The relevance of the problem of oral use of many proteins and peptides, including the GDF-11 protein, as drugs and in specialized and functional products is noted. The aim of the research is to develop a proteolysis-resistant peptide sequence of the GDF-11 protein with virtual screening of its toxicity, allergenicity, uniqueness with subsequent synthesis of the corresponding plasmid for protein expression in E. coli. The GDF-11 protein, pET-25b(+) plasmids for expression in E. coli with the peptide sequence CTVDСFFECAFGСWDС have been used as objects of study. The GDF-11 protein gene with the sequence CTVDСFFECAFGСWDС has been synthesized from oligonucleotides using the cyclic assembly method. The obtained plasmid sequence encoding the new GDF-11 protein has been treated with restriction endonucleases BamHI and XhoI and cloned into the pET-25b(+) vector for E. coli expression. The clones have been sequenced by the Sanger method. The distribution and identification of primers in the obtained sequence have been studied by the MALDI-TOF MS Ultraflex method. A new unique peptide sequence CTVDСFFECAFGСWDС with cyclization at the C1-C4, C1-C2, C1-C3 bonds of the GDF-11 protein is developed, which allows increasing its resistance to proteolysis and, accordingly, opens up the possibility of oral use in prophylactic food products. Based on virtual screening, it is established that the new peptide chain CTVDСFFECAFGСWDС of the GDF-11 protein is not toxic to the cardiovascular and nervous systems, is not hepatotoxic, does not have cytotoxicity, does not have allergenicity and is characterized by high plasma clearance and an average half-life.https://vestnik.mauniver.ru/show-eng.shtml?art=2271proteinpeptide sequencehepatoxicitycytotoxicityallergenicityproteolysisбелокпептидная последовательностьгепатотоксичностьцитотоксичностьаллергенностьпротеолиз |
| spellingShingle | Valieva Sh. S. Tikhonov S. L. Tikhonova N. V. Timofeeva M. S. Shikhalev S. V. Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli Vestnik MGTU protein peptide sequence hepatoxicity cytotoxicity allergenicity proteolysis белок пептидная последовательность гепатотоксичность цитотоксичность аллергенность протеолиз |
| title | Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli |
| title_full | Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli |
| title_fullStr | Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli |
| title_full_unstemmed | Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli |
| title_short | Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli |
| title_sort | construction of a proteolysis resistant peptide sequence of the food recombinant protein gdf 11 and plasmid synthesis for its expression in e coli |
| topic | protein peptide sequence hepatoxicity cytotoxicity allergenicity proteolysis белок пептидная последовательность гепатотоксичность цитотоксичность аллергенность протеолиз |
| url | https://vestnik.mauniver.ru/show-eng.shtml?art=2271 |
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